1. Field of the Invention
The present inventions relates to human tachykinins, their precursors and their production by recombinant DNA technology.
The abbreviations used herein are as follows:
b--bovine PA1 h--human PA1 PPT--preprotachykinin PA1 EDTA--ethylene diamine tetracetic acid PA1 BSA--bovine serum albumin PA1 PVP--polyvinyl pyrrolidone PA1 SDS--sodium dodecyl sulphate PA1 SSC--saline sodium citrate.
2. Description of Related Art Including Information Disclosed under 35 C.F.R. .sctn..sctn.1.97-1.99.
The tachykinins are a family of small peptides, present in the brain and peripheral tissues, which play important roles as neurotransmitters and as hormones. The tachykinins are characterised by a common C-terminal amino acid sequence, -Phe-X-Gly-Leu-Met-NH.sub.2, wherein X is a hydrophobic or aromatic residue [Erspamer, V., Trends in Neuroscience, 4, 267-269, 1981].
The first tachykinin found in mammals is known as substance P, which is though to be involved in the transmission of painful stimuli in the spinal cord. It is also released from sensory nerves in the skin and is thought to play a role in the inflammatory response.
Subsequently, other tachykinins, known as neurokinin A and neurokinin B were reported [Kimura, S., Oada, M., Sugita, Y., Kanazawa, I. and Munekata, E., Proc Jap. Acad. Series B, 59, 101-104, 1983], and it has recently been shown [Tatemoto, K., Lundberg, J. M., Jornvall, H and Mutt, V., Biochem. Biophys. Res. Comm., 128, 947-953, 1985] that in the porcine central nervous system, there is a larger tachykinin, known as neuropeptide K.
It has been shown [Nawa, H., Hirose, T., Takashima, H., Inayama, S. and Nakanishi, S., Nature, 306, 32-36, 1983] that in bobine brain substance P is derived from one of two larger polypeptide precursors, known as alpha- and beta-bovine preprotachykinin (bPPT). It has been shown that the mRNAs encoding alpha- and beta-bPPTs are derived from a single bPPT gene as a result of tissue specific RNA splicing [Nawa, H., Kotani, H and Nakanishi, S., Nature 312, 729-734, 1984]. It has been shown that bovine neurokinin A is derived from beta-bPPT but cannot be derived from alpha-bPPT.
Thus, it has been shown that residues 58 to 68 and 98 to 107 of beta-bPPT correspond to bovine substance P and bovine neurokinin A. Moreover it has been shown that residues 72 to 107 correspond to the sequence of bovine neuropeptide K.